Difference between revisions of "Part:BBa K3183203"
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mClover3 is a 26.9 kDa protein derived from GFP. mClover3 improves photostability by 60% (t<sub>1/2</sub> = 80s) to its predecessor, owing to 2 mutations relative to dClover2: A206K and S160C. mClover3 can be used in Förster Resonance Energy Transfer (FRET) experiments with mRuby3, providing an alternative to cyan/yellow partners. This has the advantage of reducing spectral separation, having lower phototoxicity, and lower autofluorescence<sup>1</sup>. | mClover3 is a 26.9 kDa protein derived from GFP. mClover3 improves photostability by 60% (t<sub>1/2</sub> = 80s) to its predecessor, owing to 2 mutations relative to dClover2: A206K and S160C. mClover3 can be used in Förster Resonance Energy Transfer (FRET) experiments with mRuby3, providing an alternative to cyan/yellow partners. This has the advantage of reducing spectral separation, having lower phototoxicity, and lower autofluorescence<sup>1</sup>. | ||
| − | This construct was used for Spy&Go purification as a part of our characterisation of SpyTag, [https://parts.igem.org/Part:BBa_K1159201 BBa_K1159201.] Expression of this part was achieved in the pET28A vector [https://parts.igem.org/Part:BBa_K2141000 BBa_K2141000.] | + | This construct was used for Spy&Go purification as a part of our characterisation of SpyTag, [https://parts.igem.org/Part:BBa_K1159201 BBa_K1159201.] Expression of this part was achieved in the pET28A vector [https://parts.igem.org/Part:BBa_K2141000 BBa_K2141000.]<br><br> |
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Revision as of 09:57, 20 October 2019
mClover3 with SpyTag and 6-His for E. coli
mClover3 is a 26.9 kDa protein derived from GFP. mClover3 improves photostability by 60% (t1/2 = 80s) to its predecessor, owing to 2 mutations relative to dClover2: A206K and S160C. mClover3 can be used in Förster Resonance Energy Transfer (FRET) experiments with mRuby3, providing an alternative to cyan/yellow partners. This has the advantage of reducing spectral separation, having lower phototoxicity, and lower autofluorescence1.
This construct was used for Spy&Go purification as a part of our characterisation of SpyTag, BBa_K1159201. Expression of this part was achieved in the pET28A vector BBa_K2141000.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Characterisation
References
1) Bajar, Bryce T et al. “Improving brightness and photostability of green and red fluorescent proteins for live cell imaging and FRET reporting.” Scientific reports vol. 6 20889. 16 Feb. 2016, doi:10.1038/srep20889